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PMID: 8094885 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Modulation of glutamine synthetase gene expression in tobacco by the introduction of an alfalfa glutamine synthetase gene in sense and antisense orientation: molecular and biochemical analysis.

Molecular & general genetics : MGG ·Vol. 236 ·No. 2-3 ·1993-01-00 ·Pages 315-25

Temple SJ, Knight TJ, Unkefer PJ, Sengupta-Gopalan C

Abstract

A glutamine synthetase (GS) cDNA isolated from an alfalfa cell culture cDNA library was found to represent a cytoplasmic GS. The full-length alfalfa GS1 coding sequence, in both sense and antisense orientation and under the transcriptional control of the cauliflower mosaic virus 35S promoter, was introduced into tobacco. Leaves of tobacco plants transformed with the sense construct contained greatly elevated levels of GS transcript and GS polypeptide which assembled into active enzyme. Leaves of the plants transformed with the antisense GS1 construct showed a significant decrease in the level of both GS1 and GS2 polypeptides and GS activity, but did not show any significant decrease in the level of endogenous GS mRNA. We have proposed that antisense inhibition using a heterologous antisense GS RNA occurs at the level of translation. Our results also suggest that the post-translational assembly of GS subunits into a holoenzyme requires an additional factor(s) and is under regulatory control.

MeSH Terms
Alcohol Oxidoreductases/biosynthesis Cells, Cultured Cloning, Molecular Down-Regulation Gene Expression Genetic Variation Glutamate-Ammonia Ligase/biosynthesis,genetics Hydroxypyruvate Reductase Medicago sativa/enzymology,genetics Nucleic Acid Hybridization Phosphoenolpyruvate Carboxylase/metabolism Plant Proteins/metabolism Plants, Genetically Modified/enzymology,genetics Plants, Toxic Protein Conformation RNA, Antisense/genetics RNA, Messenger/analysis Species Specificity Tobacco/enzymology,genetics Transcription, Genetic Transformation, Genetic
Chemicals
Plant Proteins RNA, Antisense RNA, Messenger Alcohol Oxidoreductases Hydroxypyruvate Reductase Phosphoenolpyruvate Carboxylase Glutamate-Ammonia Ligase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Temple S J
Plant Genetic Engineering Labs/Department of Agronomy and Horticulture, New Mexico State University, Las Cruces 88003.
Knight T J
Unkefer P J
Sengupta-Gopalan C
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00277128
Published
1993-01-00
Pages
315-25
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00277128
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